1. Field of the Invention
The present invention relates to a process for removing an undesired component from a bound desired component, in particular for improving the bound-free separation efficiency. In particular, the present invention relates to a process for separating or washing a bound analyte being analyzed in an automated clinical analyzer from unbound label, particularly without decreasing processing efficiency or speed.
2. Description of the Related Art
Methods and systems for washing containers that hold samples being analyzed, such as analyzers for conducting clinical assays are known, e.g., wash stations in clinical analyzer immunochemical assay systems. For example, U.S. Pat. No. 6,096,561 and U.S. application Ser. No. 09/482,599 filed Jan. 13, 2000 entitled “Failure Detection in Automatic Clinical Analyzers” describe immunoassay analyzers that include container wash stations for washing containers containing one or more analytes bound to coated sample containers that are measured, for example, by chemiluminescence. Such systems typically contain a sample wash station that may include a wash fluid dispense nozzle and an aspirating nozzle. The sample containing analyte and reagent, e.g., label, is aspirated out of the container after it has been incubated. Wash fluid is then dispensed into and aspirated out of the container one or more times to remove any excess analyte and reagent not bound to the coating, such as streptavidin, at the side of the container. Some known surface coated containers have features, such as pockets or ledges, near the top of the container that can trap unbound material, such as unbound label, analyte, etc. These features may be the result of the process used to mold the containers and/or to keep the containers separated in a stack. See, e.g., U.S. Pat. No. 5,441,895, which describes stackable containers. The incubation, reagent metering and mixing processes involved in immunochemical assay analysis move the sample in the container in a manner that leaves a film of sample containing unbound label and/or analyte on the pockets and ledges that are in the upper regions of the container.
Typically in immunochemical assay systems, an important aspect that affects the performance is bound-free separation. The bound-free separation is controlled by two primary factors:                (1) the component or material, such as the bound label, that is intended to produce signal remains behind and intact; and        (2) the unbound (i.e., free) component or material is removed as completely as possible.        
In particular, there are several assays that have clinically significant performance very close to the background of the assay. This means that small amounts of unbound material present during the measurement portion of the process, particularly signal producing material, can produce a substantial adverse impact on performance.
To remove unbound material as completely as possible, a container wash process typically includes multiple wash cycles, such as filling a surface coated container to a first height on the container with a wash fluid and aspirating the wash fluid after a predetermined amount of time. For example, in one known process, a dispensing nozzle fills the container, which has a 300 μl capacity up to a height of 270 μl and then sets the soak height to 230 μl so the well can be transported during the wash incubation step (≅37-40 seconds). These steps, including the incubation step, are repeated multiple times (e.g., four times) using the same fluid heights. Unbound material that can be present in the upper regions of the container is only removed by inadvertent exposure to the wash fluid. Considerable erroneous signal can be generated when the wash fluid makes contact with unbound material in the upper regions of the container where the material is re-hydrated but not removed. The unbound material can then drop into the signal reagent during the last processing step that is intended to detect the amount of label bound to the container surface.
Another problem with known systems that remove fluid by aspiration is that the outer surface of an aspirating nozzle can become contaminated with the wash fluid containing unbound material. This can lead to contamination of the container with unbound material in subsequent wash cycles (in that particular test or across subsequent tests). Still another problem with known systems that unbound material can reside or float on top of the fluid, even after several wash steps. This can also interfere with any subsequent analysis of the desired bound material. It is believed that surfactant in wash fluid can trap unbound material enabling it to float. Upon the addition of an additional fluid, such as a signal reagent, the unbound material, can float to the surface of the fluid, where it will interfere with subsequent analysis. This may particularly be the case if the unbound material is horseradish peroxidase (“HRP”), which when bound to denatured protein may also prefer to float on the surface of the signal reagent fluid, where it will combine with the signal associated with bound HRP and create a false signal which will be read, thereby leading to erroneous results.
The problems described above may lead to the unbound material remaining in the test container and possibly interfering with the subsequent analysis of the analyte, leading to tests that need to be repeated at considerable inconvenience and expense due to inaccurate results.
It is known in the art that the unbound material in the upper regions of the container can be removed more completely if the fluid were to be filled higher in the container and remain there for the soak cycle (i.e., container wash incubation). This is not practical for a random access analyzer system since the test element needs to be transported during the soak cycle so other tests elements can be processed. Batch analyzers leave the test element static during this process step, which allows the fluid level moved to the very top of the container (positive meniscus). Even analyzers that fill the test element to the very top of the container may still have issues with not completely removing unbound label at the very top, if there are features in this region that can trap or retain unbound material. This is especially true when each wash processing cycle raises the fluid to the same height. In this process, the last processing step can cause any residual unbound material to flow into a region of the test element where it can interact with the signal generating reagents, thus producing erroneous results.